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Statements

Subject Item
n2:RIV%2F61989592%3A15310%2F12%3A33142716%21RIV13-MSM-15310___
rdf:type
n6:Vysledek skos:Concept
rdfs:seeAlso
http://www.sciencedirect.com/science/article/pii/S0003267012012792
dcterms:description
A capillary zone electrophoresis (CZE) method for separation of adenosine and N-6-isopentenyladenosine (cytokinin) nucleotides was developed, optimized and validated. Aqueous solutions of several amino acids were evaluated as the background electrolyte constituents. Separation of six nucleotides in less than 20 min with high theoretical plate number (up to 400 000 for isopentenyladenosine triphosphate) was achieved using a 100 mM sarcosine/ammonia buffer at pH 10.0. The detection limits of the CZE-UV method are in the low micromolar range (0.69-1.27 mu mol L-1). Good repeatability of migration times (within 1.3%), peak areas (within 1.8%) and linearity (R-2 }0.999) was achieved over the concentration range 5-1000 mu mol L-1. The method was used to assay the activity of the recombinant Arabidopsis thaliana isopentenyltransferase 1 (AtIPT1). Baseline separation of isopentenylated nucleotides by CE-ESI-MS using a volatile buffer (30 mM ammonium formate; pH 10.0) was accomplished. The identities of the reaction products - isopentenyladenosine di- and triphosphate were confirmed by HPLC-QqTOE-MS. Dephosphorylation of ATP was observed as a parallel reaction A capillary zone electrophoresis (CZE) method for separation of adenosine and N-6-isopentenyladenosine (cytokinin) nucleotides was developed, optimized and validated. Aqueous solutions of several amino acids were evaluated as the background electrolyte constituents. Separation of six nucleotides in less than 20 min with high theoretical plate number (up to 400 000 for isopentenyladenosine triphosphate) was achieved using a 100 mM sarcosine/ammonia buffer at pH 10.0. The detection limits of the CZE-UV method are in the low micromolar range (0.69-1.27 mu mol L-1). Good repeatability of migration times (within 1.3%), peak areas (within 1.8%) and linearity (R-2 }0.999) was achieved over the concentration range 5-1000 mu mol L-1. The method was used to assay the activity of the recombinant Arabidopsis thaliana isopentenyltransferase 1 (AtIPT1). Baseline separation of isopentenylated nucleotides by CE-ESI-MS using a volatile buffer (30 mM ammonium formate; pH 10.0) was accomplished. The identities of the reaction products - isopentenyladenosine di- and triphosphate were confirmed by HPLC-QqTOE-MS. Dephosphorylation of ATP was observed as a parallel reaction
dcterms:title
Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction
skos:prefLabel
Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction Analysis of cytokinin nucleotides by capillary zone electrophoresis with diode array and mass spectrometric detection in a recombinant enzyme in vitro reaction
skos:notation
RIV/61989592:15310/12:33142716!RIV13-MSM-15310___
n6:predkladatel
n7:orjk%3A15310
n3:aktivita
n9:Z n9:P
n3:aktivity
P(ED0007/01/01), P(GA522/08/0920), P(LC06034), Z(AV0Z50380511), Z(MSM6198959216)
n3:cisloPeriodika
NOV
n3:dodaniDat
n13:2013
n3:domaciTvurceVysledku
n4:5991803 n4:4290860 n4:8479089 n4:4712048 n4:1749560 n4:2098628 n4:4866029
n3:druhVysledku
n14:J
n3:duvernostUdaju
n5:S
n3:entitaPredkladatele
n21:predkladatel
n3:idSjednocenehoVysledku
122103
n3:idVysledku
RIV/61989592:15310/12:33142716
n3:jazykVysledku
n16:eng
n3:klicovaSlova
Cytokinin biosynthesis; Isopentenyltransferase; Capillary electrophoresis; Cytokinin nucleotides
n3:klicoveSlovo
n10:Capillary%20electrophoresis n10:Cytokinin%20biosynthesis n10:Cytokinin%20nucleotides n10:Isopentenyltransferase
n3:kodStatuVydavatele
NL - Nizozemsko
n3:kontrolniKodProRIV
[981CD75F0D80]
n3:nazevZdroje
Analytica Chimica Acta
n3:obor
n15:ED
n3:pocetDomacichTvurcuVysledku
7
n3:pocetTvurcuVysledku
11
n3:projekt
n11:LC06034 n11:GA522%2F08%2F0920 n11:ED0007%2F01%2F01
n3:rokUplatneniVysledku
n13:2012
n3:svazekPeriodika
751
n3:tvurceVysledku
Doležal, Karel Strnad, Miroslav Spíchal, Lukáš Béres, Tibor Dessoy, Marco A Ganzera, Markus Friedecký, David Gemrotová, Markéta Wessjohann, Ludger A Tarkowski, Petr Maier, Vítězslav
n3:wos
000311013100021
n3:zamer
n8:MSM6198959216 n8:AV0Z50380511
s:issn
0003-2670
s:numberOfPages
6
n17:doi
10.1016/j.aca.2012.08.049
n19:organizacniJednotka
15310